Abstract
A reliable indirect competitive enzyme-linked immunosorbent assay (ELISA) based on a new specific monoclonal antibody was developed to determine olaquindox in animal feed. The influence of several physicochemical factors (nonfat dried milk solution, organic solvent, incubation time) on the immunoassay was investigated. In the optimized system, the 50% inhibition concentration was 9.66 ± 1.81 µ g L−1. The limits of detection for porcine, chicken, and fish feed were 0.28, 0.46, and 0.48 µg kg−1. The limits of quantification were 1.00 µg kg−1 for the feed samples. The recoveries from porcine, chicken, and fish feed spiked with olaquindox were 90–104%, 77–103%, and 78–107%, respectively, with coefficients of variation (CVs) between 3.8 and 14.1%. The cross-reactivity was less than 2.08% with four structurally related compounds and no recognition of five other restricted or forbidden drugs was observed. Parallel analysis of the three spiked feed samples showed comparable results between the indirect competitive ELISA and the standard high-performance liquid chromatography method in China (R2 = 0.9985 for porcine feed, R2 = 0.9896 for chicken feed, and R2 = 0.9987 for fish feed). These data suggest that the developed indirect competitive ELISA is a specific and convenient method and is suitable for olaquindox determination in animal feed.
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